For determination of the protein profiles, the testicular proteins (80 [micro]g) were loaded and separated on 10 % sodium dodecyl sulfate (SDS) polyacrylamide gel followed by Coomassie blue
Electrophoris was run at a constant voltage of 200V with gel stained for 12 hours with Coomassie Blue
Brilliant R-250 0.
staining of proteins separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).
Gels were stained with Coomassie blue
and 20 [micro]L samples were loaded perlane.
The purity of the purified protein was assessed using electrophoresis on the 12% polyacrylamide gel and subsequent Coomassie blue
We excised well resolved spots from coomassie blue
stained 2 DE gel by using Ex-Quest spot cutter (Bio-Rad).
As protein loading controls, the SDS-PAGE gel was stained with Coomassie blue
(Figure 7(b)) and the blot was probed for GAPDH (Figure 7(b)).
The gel was stained with Coomassie Blue
: The gel was incubated in Coomassie blue
After 2-DE, for MS identification, proteins in gel were stained by a modified colloidal Coomassie blue
BSA concentrations were obtained from the Bradford method relying on the binding of the dye Coomassie Blue
G250 to protein , The absorbance at 595 nm was determined by a UV-vis spectrophotometer (UV-1601, Shimadzu, Japan).
After staining the membrane with coomassie blue
, the spots were excised and the N-terminal sequence of the proteins determined using an automated Edman degradation method with a Perkin Elmer Applied Biosystems Model 494 Procise protein sequencer (Foster City, CA, USA).
Protein preparation was subjectd to electrophoresis in SDS- polyacrylamide gel (12 %) (Laemmli 1970)  and protein was visualised by Coomassie blue